Immunomodulatory Effect of Parkia Speciosa Hassk. Pods Extract on Rat

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Original Article | Artículo Original

Immunomodulatory effect of Parkia speciosa Hassk. pods extract on rat induced by Salmonella typhimurium [Efecto inmunomodulador del extracto de vainas de Parkia speciosa Hassk. en ratas inducidas por Salmonella typhimurium] Fitrya Fitrya1*, Annisa Amriani1, Rennie Puspa Novita1, Elfita2, Dewi Setiorini1

1Pharmacy Department, Sriwijaya University, Palembang, Indonesia. 2Chemistry Department, Sriwijaya University, Palembang, Indonesia. *E-mail: [email protected]

Abstract Resumen Context: Parkia speciosa, which has phenolic compounds and Contexto: Parkia speciosa, que tiene compuestos fenólicos y actividades pharmacology activities, is a plant having potential immunomodulatory farmacológicas, es una planta que tiene agentes inmunomoduladores agents from the extract of its pod. potenciales del extracto de su vaina. Aims: To evaluate the potential of P. speciosa as an immunomodulatory Objetivos: Evaluar el potencial de P. speciosa como agente agent tested on albino rats induced by the bacteria Salmonella inmunomodulador en ratas albinas inducidas por la bacteria Salmonella typhimurium leading to typhoid fever. typhimurium que conduce a la fiebre tifoidea. Methods: The animals were divided into six groups: normal non-infected Métodos: Los animales se dividieron en seis grupos: grupo normal no group, negative control group, positive control, and three test groups infectado, grupo de control negativo, control positivo y tres grupos de with doses of 200, 400 and 800 mg/kg BW. The test groups were treated prueba con dosis de 200, 400 y 800 mg/kg de peso corporal. Los grupos by the extract for 12 days. The control and test groups were induced de prueba fueron tratados con el extracto durante 12 días. El control y with the S. typhimurium on the 8th day. On day 12th, CD4+, total los grupos de prueba fueron inducidos con S. typhimurium en el octavo leukocytes, differential cell count, and histology of the spleen organ día. El día 12, se evaluaron los CD4+, los leucocitos totales, el recuento were evaluated. Statistical analysis was performed by using one-way diferencial de células y la histología del órgano del bazo. El análisis ANOVA followed post hoc LSD test (α 0.05). estadístico se realizó mediante el uso de ANOVA unidireccional seguido Results: The result showed that the ethanol extract of P. speciosa pod de la prueba post hoc LSD (α 0,05). could increase the number of CD4+. The extract to 200 and 400 mg/kg Resultados: El extracto etanólico de la vaina de P. speciosa podría BW doses increased in the parameters of leukocyte, neutrophil, aumentar el número de CD4+. El extracto a dosis de 200 y 400 mg/kg de lymphocyte and monocyte with insignificant differences compared to peso corporal aumentó en los parámetros de leucocitos, neutrófilos, that of the positive control group (p>0.05). However, there were linfocitos y monocitos con diferencias insignificantes en comparación significant differences between the two first parameters between the test con el grupo de control positivo (p>0,05). Sin embargo, hubo diferencias group with 800 mg/kg BW doses compared to the positive control significativas entre los dos primeros parámetros entre el grupo de group. Moreover, more necroses in the spleen were observed from prueba con dosis de 800 mg kg de peso corporal en comparación con el histological analysis on 800 mg/kg BW doses than 400 mg/kg BW. grupo de control positivo. Además, se observaron más necrosis en el Conclusions: In general, P. speciosa ethanolic extract can increase the bazo a partir del análisis histológico con dosis de 800 mg/kg de peso immune system based on the CD4+ count, leukocyte, monocyte, and corporal que con 400 mg/kg de peso corporal. neutrophil, and causes normal necroses in low doses. Conclusiones: En general, el extracto etanólico de P. speciosa puede aumentar el sistema inmunitario en función del recuento de CD4+, leucocitos, monocitos y neutrófilos, y causa necrosis normales en dosis bajas. Keywords: Parkia speciosa; immunomodulator; Salmonella typhimurium; Palabras Clave: Parkia speciosa; inmunomodulador; Salmonella typhoid fever. typhimurium; fiebre tifoidea.

ARTICLE INFO Received: March 21, 2020. Received in revised form: May 29, 2020. Accepted: May 31, 2020. Available Online: June 11, 2020. Declaration of interests: The authors declare no conflict of interest. Funding: This work was supported by funding from Research and Community Service Institution of Sriwijaya University (Indonesia) by mean the project “Hibah Penelitian Unggulan Kompetitif 2019” [No. 0149.159/UN9/SB3.LP2M.PT/2019].

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Fitrya et al. Immunomodulator effect of Parkia speciosa

INTRODUCTION ed as an antioxidant, anti-carcinogenic and anti- inflammatory (Kamisah et al., 2013; Mustafa et al., Infectious diseases are the second leading cause 2018), antianemia (Nursucihta et al., 2014), antihy- of death in worldwide after cardiovascular disease pertensive and heart problems (Siow and Gan, (WHO, 2004). Infection can be caused by viruses, 2013; Kamisah et al., 2017) and antibacterial (Uyub bacteria, protozoa, worms, and parasitic fungi en- et al., 2010). However, the immunomodulatory tering the body or growing on the body surface. activity of P. speciosa has not been found in previ- An infectious disease with a high occurrence in ous studies. Thus, this study aims to reveal the several Asian countries is typhoid fever (Purba et potential of P. speciosa as an immunomodulatory al., 2016). This acute fever is caused by bacterium agent by observing CD4+ parameters, total leuko- Salmonella enterica infection, especially its deriva- cytes, differential blood cell counts, and spleen’s tive, Salmonella typhimurium (Alba et al., 2016). evaluation of albino rats induced by Salmonella Based on WHO data, the number of typhoid fever typhimurium bacteria. patients in Indonesia is relatively high, reaching 81,000 of 100,000 inhabitants (Rahmasari and Les- MATERIAL AND METHODS tari, 2018). A person with a weak immune system is at a high risk of developing typhoid fever (Kalia Vegetal material and chemicals et al., 2016). P. speciosa was collected from Musi Rawas dis- The innate immune system, which consists of tricts, South Sumatera, Indonesia (2.90°S, phagocyte cells, is the most important to defense 103.28°E). This species has been identified by the against microorganisms and malignant cells (Ha- Indonesian Institute of Science with Register No. run et al., 2015; Venkatalakshmi et al., 2016). Acti- 218/IPH.06/HM/I/2018. Salmonella typhimurium vation of the immune system is required to help a ATCC 14028 (105 CFU) was obtained from the body destroying antigenic substances, and it can Central Laboratory of Health, Palembang, South be stimulated using immunomodulators. Sumatera, Indonesia. Chemicals as methanol, Synthetic immunomodulatory drugs have NaCl (Merck), phosphate buffer pH 6,8 and Giem- many benefits, but adverse side effects limit their sa stain (Sigma-Aldrich) were in an analytical use. Further studies are thus an important way to grade. find safer and more effective immunomodulatory Preparation of P. speciosa pod extract agents (Venkatalakshmi et al., 2016). Plant extracts are commonly considered as potential agents from Ethanol extract of P. speciosa pod was prepared their immunomodulatory properties, which have by a maceration method. Its powder (500 mg) was smaller side effects (Alamgir and Uddin, 2010). soaked with 2 L of ethanol 95% for 2 days. A re- Their properties are generated by activating a maceration process of the residue was carried out function and efficiency of macrophages, granulo- twice. The maceration result then was evaporated cytes, complement, and natural killer cells and by with a rotary evaporator (Yamato®) at a tempera- producing effector molecules (Jayathirtha and ture of 65°C to get a concentrated extract (Harun et Mishra, 2004). Plant metabolites such as sterols, al., 2015). polysaccharides, alkaloids, flavonoids, lectins, and glycoproteins are used as immunomodulatory Design of test animal experiments agents (Harun et al., 2015). The pod of Parkia speci- Wistar male white albino rats (150 - 200 g osa Hassk. (Leguminosae) contains phytochemical weight) were used in this study. An ethics certifi- compounds such as flavonoids, phenolics, alka- cate for the use of test animals was obtained from loids, and saponins (Kamisah et al., 2013). Phar- the Faculty of Medicine, Sriwijaya University, with macology activities of P. speciosa have been report- No. 064/kepkrsmhfkunsri/2019. The test animals http://jppres.com/jppres J Pharm Pharmacogn Res (2020) 8(5): 458

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were acclimatized to the laboratory environment cells were manually analyzed by peripheral blood at a temperature of 23 ± 2°C and a 12-hour light- smear observation under a microscope (Olympus dark cycle for one week. During the acclimation CX21®). process, these rats were given standard ad libitium drink and feed (Johnson et al., 2017). They were Evaluation of spleen organs divided into 6 groups: the normal group and nega- Peritoneal and thoracic cavities were opened af- tive control group were given Tween-80 0.5 ter euthanizing by cervical dislocation. Spleen or- mL/kg BW; the positive control group was given gans were isolated and washed with distilled water 0.54 mL (dose 2.5 mg/kg BW) of Stimuno® syrup and 0.9% NaCl, consecutively. A macroscopic ex- (containing extracts of Centella asiatica, 5 mg/mL); amination was observed based on size, weight, the test groups I-III received 200, 400 and 800 color, and consistency of spleen organs. Then the mg/kg BW of the extract, respectively. All groups spleen was fixed with 10% formalin for histologi- (except the normal group) were also given 0.5 mL cal analysis. Salmonella typhimurium 105 CFU (intraperitoneal) on the 8th day of treatment as the agent inducer of Statistical analysis the typhoid fever. The CD4+ and the differential counts were ana- Antigen preparation lyzed using SPSS for Windows. Macroscopic find- ings of the spleen of each test group were com- S. typhimurium bacterial culture was bred in pared to the control group. The normality test was slant Salmonella-Shigella Agar (SSA) media, and it performed by using a Shapiro-Wilk analysis fol- was then incubated at a temperature of 37°C for 48 lowed by one way ANOVA. A post hoc LSD test h. The bacterial culture was suspended with 10 mL was performed to determine the significance be- of 0.9% NaCl solution in a tube until the concen- tween groups with α 0.05. tration reached 105 CFU/mL, which was measured using a densitometer. RESULTS

Immunomodulatory effects test Evaluation of CD4+ counts After the acclimatization period, the test ani- The result of the CD4+ count measurement mals were orally treated with the extract once a showed in Table 1. ANOVA test showed that the day. The animals were induced by giving antigen ethanol extract of P. speciosa pod could increase the suspension of S. typhimurium dose of 105 CFU as number of CD4+ compared to those of the normal much as 0.5 mL on the 8th day. Before and 24 h group (p<0.05). Administration of the ethanol ex- after the induction, the body temperature of the tract of P. speciosa pod in the doses of 400 and 800 albino rats was measured. Furthermore, blood was mg/kg body weight were sufficient doses to in- taken, and spleen was observed on the 12th day crease CD4+ significantly. (Susanti et al., 2012). Differential blood cell evaluation Evaluation of leukocytes and differential cells The statistical analysis results of leucocyte and Test animals were intraperitoneally anesthe- differential blood cell measurements of three test tized with phenobarbital 40 mg/kg BW. By using groups (I, II, and III), and the control groups (nor- cardiac puncture, the blood sample was collected mal, positive, and negative) based on the signifi- as much as 1 mL to transfer it into the EDTA vacu- cant values are presented in Table 2. Based on tainer tube. Then, the total determination of leuko- these results, even though there are statistically cytes was counted by using a hematology analyzer significant differences in leukocyte and lympho- (Sysmex® KX-21). Meanwhile, CD4+ counts were cyte counts between all test groups and two con- analyzed using a flow cytometer (Alere® Pima trol (normal and negative) groups, differences be- analyzer) (Yapo et al., 2011). The differential blood http://jppres.com/jppres J Pharm Pharmacogn Res (2020) 8(5): 459

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tween the two first test groups and the positive Based on the histological observations, levels of group are insignificant. In addition, the significant necrosis occurred in the spleens of the positive value of the difference between the test group III control group and the test group were lower than and the positive one was less than 0.05. These val- those of the negative control group. The lowest ues of the significant differences imply that the level of necrosis was found in the group I and the leukocyte counts can be stimulated in all test positive group. The results of this histological and groups; however, the group III can increase the macroscopic observation of the spleens are in good leukocyte counts higher than the groups I and II. agreement, which showed group I exhibited a weight, color, and consistency relatively similar to Furthermore, the statistically significant values the normal group. of differences between the test groups (I and II) and the positive control group were higher than Fig. 2A shows that in the spleen histology of the 0.05 for two parameters of the differential blood normal group, many follicles were found, but no cell, i.e., lymphocyte and monocyte. Therefore, necrosis was observed. Otherwise, the most severe they did not show statistically significant differ- necrosis occurred in the negative control group ences in these parameters. Contrary, the compari- compared to other groups (Fig. 2C). Like the posi- son between test group III and the positive group tive control group (Fig. 2B), the test rats treated showed statistically significant differences (p<0.05) with the extract of P. speciosa pod at the lowest for the last parameter (neutrophils). dose, 200 mg/kg BW (Fig. 2D), had the least necrosis. Therefore, it indicates that the extract can Spleen evaluation reduce necrosis as observed in the test group as The shape of a normal spleen is likely an elon- well as in the positive control group. The lower gated bean with the sharp in one of the edges and doses the extract was given, the less necrosis oc- red to dark red. Damaged spleen will be blackish, curred, and vice versa. and the edges will tend to be rounded or blunt Moreover, at 400 and 800 mg/kg BW doses of (Cesta, 2006). The macroscopic spleen of rats can the extract, a considerable amount of the necrosis be presented in Fig. 1. It can be clearly seen that was observed (Fig. 2E-F). It may suggest that the the spleen of the negative group has a black-bigger higher the dose was given, the more accumulation shape and harder consistency than that of other of the lymphocyte and macrophage observed in groups. The statistical analysis showed that group the spleen, and as a consequence, the effect of hy- III had significantly heavier spleens compared to perplasia is higher. the positive control group (p<0.05) (Table 3).

Table 1. CD4+ count in rats with typhoid fever and treated with P. speciosa pod ethanolic extract. Group Treatment Dose CD4+ (cells/mm3)

Normal Tween-80 0.5 mL/kg 6.0 ± 0.00 Negative control Tween-80 0.5 mL/kg 4.0 ± 0.00 Positive control Centella asiatica 2.5 mg/kg 6.5 ± 0.70 Group I P. speciosa 200 mg/kg 7.5 ± 4.95a

Group II P. speciosa 400 mg/kg 8.5 ± 3.54ab

Group III P. speciosa 800 mg/kg 11.0 ± 7.07ab

Values are presented as mean ± SD, n=3; ap<0.05 statistically significant differences compared with the positive control group: group I (p=0.016), group II (p=0.001), and group III (p=0.00); bp<0.05 statistically significant differences compared with the normal group with p values for group II (p=0.023), and group III (p=0.00). Salmonella typhimurium 0.5 mL (105 CFU, intraperitoneal) was administered on the 8th day of treatment in all groups except normal group.

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Table 2. Differential blood cell counts in animals with typhoid fever and treated with P. speciosa pod ethanolic extract.

Group Leukocyte Lymphocyte Monocyte Neutrophils (103 /µL) (%) (%) (%)

Normal 7.83 ± 0.52 78.00 ± 3.60 11.67 ± 2.51 10.33 ± 2.88 Negative control 11.17 ± 2.14 76.00 ± 2.00 9.67 ± 1.52 10.00 ± 1.00 Positive control 14.83 ± 5.51 65.00 ± 3.00 13.00 ± 1.73 16.67 ± 1.53 Group I 15.53 ± 7.30ab 69.00 ± 1.00ab 11.67 ± 0.57b 15.33 ± 0.58b

Group II 16.57 ± 0.20ab 67.33 ± 1.15ab 15.00 ± 1.00b 20.00 ± 1.00ab

Group III 19.30 ± 5.54ac 66.00 ± 3.60ab 18.33 ± 0.57ab 24.00 ± 1.00ac

The values are presented as mean ± SD, n=3; ap<0.05 compared to the normal group; bp>0.05 compared to the positive control group; cp<0.05 compared to the positive control group. There were statistically significant differences between leucocyte (all group: p=0.00), lymphocyte (group I, p=0.006; group II, p=0.002; group III, p=0.001), monocyte (group III, p=0.024) and neutrophils (group II, p=0.03; group III, p=0.00) compare to the normal group. There were no statistically significant differences between in leukocyte (group I, p=0.67; group II, p=0.121); lymphocyte (group I, p=0.177; group II, p=0.419; group III, p=0.726); monocyte (group I, p=0.646; group II, p=0.493; group III, p=0.084); neutrophils (group I, p=0.626; group II, p=0.235) compare to positive control group but there were statistically significant differences between leukocyte (group III, p=0.00) and neutrophils (p=0.018) compare to the positive control group. Normal and negative control groups: Tween-80 0.5 mL/kg BW; positive control group: Centella asiatica (2.5 mg/kg BW); groups I-III: P. speciosa pod ethanolic extract at doses of 200, 400 and 800 mg/kg BW. Salmonella typhimurium 0.5 mL (105 CFU, intraperitoneal) was administered on the 8th day of treatment in all groups except normal group.

Table 3. Spleen morphology of the animals with typhoid fever and treated with P. speciosa pod ethanolic extract.

Morphology Treatment

Normal Negative Positive control Group I Group II Group III control

Color Red Red Black Blackish red Red with little black Blackish red Blackish red

Shape Normal Normal Normal Normal Normal Normal

Weight (g) 0.54 ± 0.05 1.03 ± 0.05 0.66 ± 0.01 0.61 ± 0.02 0.75 ± 0.05 0.88 ± 0.03a

Consistency Springy Springy hard Springy little hard Springy Springy Springy

Values are presented as means ± SD, n=3. ap<0.05 compared to the positive control group. There were statistically significant differences between group I (p=0.00), group II (p=0.00), group III (p=0.017) compare to the normal group. There were no statistically significant differences between group I (p=0.089), group II (p=0.849) compare to the positive control group, but there were statistically significant differences with the group III (p=0.017). Normal and Negative control groups: Tween-80 0.5 mL/kg BW; Positive control group: Centella asiatica (2.5 mg/kg BW); groups I-III: P. speciosa pod ethanolic extract at doses of 200, 400 and 800 mg/kg BW. Salmonella typhimurium 0.5 mL (105 CFU, intraperitoneal) was administered on the 8th day of treatment in all groups except normal group.

A B C D E F

Figure 1. Macroscopic of spleens in normal group (A); test groups I-III (B-D); negative control (E); and positive control (F). Normal and negative control groups: Tween-80 0.5 mL/kg BW; positive control group: Centella asiatica (2.5 mg/kg BW); groups I-III: P. speciosa pod ethanolic extract at doses of 200, 400 and 800 mg/kg BW. Salmonella typhimurium 0.5 mL (105 CFU, intraperitoneal) was administered on the 8th day of treatment in all groups except normal group.

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A B

C D

E F

Figure 2. Histopatology of spleen in normal group (A), positive control (B), negative control (C), doses of P. speciose ethanolic extract 200, 400, and 800 mg/kg BW (D-F). Normal and negative control groups: Tween-80 0.5 mL/kg BW; positive control group: Centella asiatica (2.5 mg/kg BW). Salmonella typhimurium 0.5 mL (105 CFU, intraperitoneal) was administered on the 8th day of treatment in all groups except normal group. H & E staining. Magnification 400.

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DISCUSSION ity of flavonoids to stimulate lymphocyte proliferation resulted in the acceleration of a lymphocyte The CD4+ is a marker on the surface of white formation to increase the lymphocyte count. How- blood cells of humans, especially lymphocyte cells. ever, as the lymphocyte was formed too quickly, it It is well known that infection causes a decrease in would die quickly. Therefore, the total number of the immune response by blocking the activity of lymphocytes in the spleen finally decreased. An- CD4+ and CD8+ cells. CD4+ has an important role other factor was the infection, which retained in the immune system. A decrease in its level im- lymphocyte in lymphoid organs such as the plies a weak immune system for blocking infec- spleen. As a consequence, lymphocyte and macro- tion. Therefore, to provide defense against the in- phage would accumulate in the spleen, and the fection, the immune system needs to be strength- spleen then swelled. (Fahrimal et al., 2014). ened by stimulating CD4+ production (Yapo et al., 2011). The spleen is an organ where immune cells, especially phagocyte cells, carry antigens, interact In addition, the activation of immune cells such with, and then produce T lymphocyte. Thus, an as T cells of CD4+ and CD8+ is a possible mecha- increase in the spleen weight is an indicator of the nism to stimulate immune activity. An increase in stimulation in the immune system (Ketema et al., the activity of the immune cells is considered a 2015). The higher the extract dose was given, the good indicator of a better prognosis and active higher ability of the extract stimulated lymphocyte immune response to tumors and infections (Gau- and macrophage cells. As this stimulation affected tam et al., 2009; Ayeka et al., 2017). In this re- the accumulation of antigens, hyperplasia due to search, an increase in the number of CD4+ is alleg- infection took place in the spleen. This spleen pro- edly triggered by flavonoid and phenolic com- duced more lymphocyte and macrophage to deal pounds contained in the ethanol extract of Parkia with antigen attacks (Cesta, 2006; Vásquez et al., speciosa. Flavonoid compounds activate T cells to 2015). differentiate into CD8+ or CD4+ cells. The CD4+ cells have no direct killing activity in infected cells CONCLUSIONS but direct other immune cells to act against cells infected with pathogens, mainly by secreting some This study proved that the ethanol extract of cytokines (Grigore, 2017). Parkia speciosa pod increases the CD4+ count and the differential cells so that it is potential as an + The levels of the CD4 cells, leukocyte, mono- immunomodulatory agent. However, an increase cyte, and neutrophil in the blood circulation can in its doses caused a decrease in lymphocyte increase due to the activation of immune- count. In addition, necrosis and hyperplasia oc- promoting compounds. The extract-contained fla- curred in the spleen. The extract with 200 mg/kg vonoid stimulates the proliferation of the number BW dose can stimulate the immune system even of differential T and B lymphocyte (Ketema et al., though it creates low necrosis. 2015). This flavonoid increases the secretion of cytokine interleukin-2 (IL-2), acting as prolifera- CONFLICT OF INTEREST tion and differentiation factors (Middleton et al., 2000). Meanwhile, phenolic compounds can stimu- The authors declare no conflict of interest. late the immune system from its hydroxyl groups. ACKNOWLEDGMENTS These compounds influence enzymes or electron transfer systems to produce immunomodulatory The authors would like to thank you to the Research and properties, especially a phagocytic activity Community Service Institution of Sriwijaya University (Indo- (Manosroi et al., 2003). nesia) for research funding through “Hibah Penelitian Unggu- lan Kompetitif 2019” [No. 0149.159/UN9/SB3.LP2M.PT- The decrease in the lymphocyte count in this /2019]. study was affected by two factors. Firstly, the abil- http://jppres.com/jppres J Pharm Pharmacogn Res (2020) 8(5): 463

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REFERENCES damage in rats given N(G)-nitro-L-arginine methyl ester. Biomed Pharmacother 96: 291–298. Alamgir M, Uddin SJ (2010) Recent advances on the Ketema T, Yohannes M, Alemayehu E, Ambelu A (2015) ethnomedicinal plants as immunomodulatory agents in Evaluation of immunomodulatory activities of Chattopadhyay D (ed.) Ethnomedicine: A Source of methanolic extract of khat (Forsk) and cathinone in Swiss Complementary Therapeutics. Kerala, India. pp. 227–244. albino mice. BMC Immunol 16(1): 1–11. Alba S, Bakker MI, Hatta M, Scheelbeek PFD, Dwiyanti R, Manosroi A, Saraphanchotiwitthaya A, Manosroi J (2003) Usman R, Sultan AR, Sabir M, Tandirogang N, Amir M, Immunomodulatory activities of Clausena excavata Burm. Yasir Y, Pastoor R, Van BS, Smits HL (2016) Risk factors wood extracts. J Ethnopharmacol 89: 155–160. of typhoid infection in the Indonesian archipelago. PLoS ONE 11(6): 1–14. Middleton E, Kandaswami C, Theoharides TC (2000) The effects of plant flavonoids on mammalian cells: Ayeka PA, Bian YH, Githaiga PM, Zhao (2017) The Implications for inflammation, heart disease, and cancer. immunomodulatory activities of licorice polysaccharides Pharmacol Rev 52(4): 673–751. (Glycyrrhiza uralensis Fisch.) in CT 26 tumor-bearing mice. BMC Complement Alternat Med 17(1): 1–9. Mustafa NH, Ugusman A, Jalil J, Kamisah Y (2018) Anti- inflammatory property of Parkia speciosa empty pod Cesta MF (2006) Normal structure, function, and histology of extract in human umbilical vein endothelial cells. J the spleen. Toxicol Pathol 34(5): 455–465. Applied Pharm Sci 8(1): 152–158. Fahrimal Y, Eliawardani, Rafina A, Azhar A, Asmilia N (2014) Nursucihta S, Thai'in HA, Putri DM, Utami DN, Ghani AP Blood profile of rats (Rattus norvegicus) infected with (2014) Antianemia activity of Parkia speciosa Hassk seed Trypanosoma evansi treated with willow tree bark extract ethanolic extract. Trad Med J 19: 49–54. (Salix tetrasperma Roxb). J Kedokteran Hewan 8(2): 164– 168. Purba EI, Wandra T, Nugrahini N, Nawawi St, Kandun N (2016) Typhoid fever control program in Indonesia: Gautam M, Saha S, Bani S, Kaul A, Mishra S, Patil D, Satti NK, Challenges and opportunities [Indonesian]. Media Suri KA, Gairola S, Suresh K, Jadhav S, Qazi GN, Libangkes 26(2): 99–108. Patwardhan B (2009) Immunomodulatory activity of Asparagus racemosus on systemic Th1/Th2 immunity: Rahmasari V, Lestari K (2018) Manajemen terapi demam Implications for immunoadjuvant potential. J tifoid: Kajian terapi farmakologis dan non farmakologis. Ethnopharmacol 121(2): 241–247. Farmaka 16(1):184–194. Grigore A (2017) Plant phenolic compounds as Siow HL, Gan CY (2013) Extraction of antioxidative and immunomodulatory agents. In: Phenolic Compounds - antihypertensive bioactive peptides from P. speciosa Biological Activity. Ed. Soto-Hernandez M, Palma- seeds. Food Chem 141: 3435–3442. Tenango M, del Rosario Garcia-Mateos M. IntechOpen. Susanti R, Yuniastuti A, Iswari RS (2012) Aktivitas reactive pp. 75–98. oxigen species macrofag akibat stimulasi gel lidah buaya Harun N, Septama A, Jantan I (2015) Immunomodulatory pada infeksi Salmonella typhimurium. J MIPA 35(1): 1–10. effects of selected Malaysian plants on the CD18/11a Uyub AM, Azlan A, Fariza SS, Nwachukwu IN (2010) In-vitro expression and phagocytosis activities of leukocytes. antibacterial activity and cytotoxicity of selected Asian Pac J of Trop Biomed 5(1): 48–53. medicinal plant extracts from Penang Island Malaysia on Jayathirtha MG, Mishra SH (2004) Preliminary some pathogenic bacteria. Ethnobot Res Appl 8: 95–106. immunomodulatory activities of methanol extracts of Vásquez B, Sandoval C, Smith RL, Sol MD (2015) Effects of Eclipta alba and Centella asiatica. Phytomedicine 11(4): 361– early and late adverse experiences on morpho- 365. quantitative characteristics of Sprague-Dawley rat spleen Johnson J, Ekpo G, Ugwuoke J (2017) Immunomodulatory subjected to stress during adulthood. Int J Clin Exp potentials of ethanolic leaf extract of Phyllantus amarus in Pathol 8(4): 3624-3635. Wistar rats. Pharm Chem J 4(5): 83–88. Venkatalakshmi P, Vadivel V, Brindha P (2016) Role of Kalia P, Kumar NR, Harjai K (2016) Effect of propolis extract phytochemicals as immunomodulatory agents: A review. on hematotoxicity and histological changes induced by Int J Green Pharm 10(1): 1–18. Salmonella enterica serovar typhimurium in BALB/c mice. WHO (2004) The global burden of disease 2004. Update. Arch Biol Sci 68(2): 385–391. World Health Organization. Switzerland: WHO Press. Kamisah Y, Othman F, Qodriyah HMS, Jaarin K (2013) Parkia Yapo FA, Yapi FH, Ahiboh H, Hauhouot-Attounbre ML, speciosa Hassk.: A potential phytomedicine. Evid Based Guédé NZ, Djaman JA, Monnet D (2011) Complement Altern Med 2013: ID Article 709028. Immunomodulatory effect of the aqueous extract of Kamisah Y, Zuhair JSF, Juliana AH, Jaarin K (2017) Parkia Erigeron floribundus (Kunth) Sch beep (Asteraceae) leaf in speciosa empty pod prevents hypertension and cardiac rabbits. Trop J Pharm Res 10(2): 187–193. ______

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AUTHOR CONTRIBUTION:

Contribution Fitrya F Amriani A Novita RP Elfita Setiorini D

Concepts or ideas x x

Design x

Definition of intellectual content x x x x

Literature search x x x x x

Experimental studies x x x x x

Data acquisition x x x x x

Data analysis x x x x x

Statistical analysis x x

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Citation Format: Fitrya F, Amriani A, Novita RP, Elfita, Setiorini D (2020) Immunomodulatory effect of Parkia speciosa Hassk. pods extract on rat induced by Salmonella typhimurium. J Pharm Pharmacogn Res 8(5): 457–465.

http://jppres.com/jppres J Pharm Pharmacogn Res (2020) 8(5): 465